Discipline of Ophthalmology & Visual Sciences, Level 7 Adelaide Health and Medical Sciences Building, University of Adelaide, North Terrace, Adelaide, SA, 5000, Australia.
South Australian Institute of Ophthalmology, Royal Adelaide Hospital, Port Road, Adelaide, SA, 5000, Australia.
Eye (Lond). 2023 Jan;37(1):62-68. doi: 10.1038/s41433-021-01872-9. Epub 2022 Jan 10.
To establish cultures of human lacrimal gland from patient-derived, biopsy-sized, tissue specimens.
Tissue was obtained after surgical removal from patients without dry eye disease undergoing routine procedures. Samples were subjected to mechanical and enzymatic digestion and resulting cell suspensions were plated onto collagen-coated glass coverslips and grown for up to 21 days. Cultures were analysed by immunocytochemistry and light microscopy, and resultant cellular distributions were compared to those in sections of fixed human lacrimal gland tissue.
Dissociation of biopsy-sized pieces of human lacrimal gland and seeding onto coated surfaces allowed development of a mixed population of cells in vitro. Within 7-14 days, cellular aggregation was observed and by 21 days many cells had organised themselves into distinct three-dimensional complexes. Immunohistochemistry revealed a heterogeneous population of cells, including epithelial, myoepithelial, mesenchymal and progenitor cells. Some of the epithelia labelled positively for lysozyme and lactoferrin.
Collection and dissociation of biopsy-sized pieces of human lacrimal gland leads to a cellular preparation that can proliferate in vitro and organise into three-dimensional structures. This is the first report detailing that biopsy-collected specimens of human lacrimal gland can be used to establish cell cultures.
从患者来源的活检大小组织标本中建立人泪腺的培养物。
在没有干眼症的患者接受常规手术切除后获得组织。对样本进行机械和酶消化，将得到的细胞悬液接种到涂有胶原蛋白的玻璃盖玻片上，并培养长达 21 天。通过免疫细胞化学和光学显微镜分析培养物，并将所得细胞分布与固定的人泪腺组织切片进行比较。
将活检大小的人泪腺组织分离并接种到涂覆的表面上，允许在体外开发出混合细胞群体。在 7-14 天内，观察到细胞聚集，在 21 天内，许多细胞已经组织成独特的三维复合物。免疫组织化学显示出异质细胞群体，包括上皮细胞、肌上皮细胞、间充质细胞和祖细胞。一些上皮细胞对溶菌酶和乳铁蛋白呈阳性标记。
收集和分离活检大小的人泪腺组织会导致可在体外增殖并组织成三维结构的细胞制剂。这是首次详细报道活检收集的人泪腺标本可用于建立细胞培养物。
